White pulp compartments in the spleen of the turtle Mauremys caspica

Summary The aim of the present study was to analyze the nature of lymphoid and non-lymphoid cellular components occurring in distinct histological compartments of the splenic white pulp of the turtle, Mauremys caspica, in order to define their possible correlations with those of the spleen of higher vertebrates, principally mammals. The white pulp of M.caspica consisted of 3 clearly distinguishable regions: (1) the periateriolar lymphoid sheath, and (2) the inner and (3) the outer zones of the periellipsoidal lymphoid sheath. Reticular cells intimately associated with reticular fibres constituted an extensive meshwork in the periarteriolar lymphoid sheath which housed principally Ig-negative lyphoid cells, mature and immature plasma cells, and interdigitating cells. A few Ig-positive cells were also present in the peripheral region of the periarteriolar lymphoid sheath. The inner and outer zones of the periellipsoidal lymphoid sheath were separated by a discontinuous layer of reticular cell processes. In the inner zone, surface Ig-positive lymphoid cells predominated as well as dendritic cells, resembling ultrastructurally the mammalian follicular dendritic cells, although no germinal centres were found in the turtle spleen. Macrophages, some cytoplasmic Ig-positive cells, and Ig-negative lymphoid cells appeared in the outer zone of the periellipsoidal lymphoid sheath. These results allow us to speculate on a phylogenetic relationship between the periarteriolar lymphoid sheath and the inner and the outer zones of the periellipsoidal lymphoid sheath of the spleen of M. caspica and the periarteriolar lymphoid tissue, the lymphoid follicles and the marginal zone, respectively, of the mammalian splenic white pulp.     

Interdigitating cells in the thymus of the turtle Mauremys caspica

Summary Interdigitating cells are non-lymphoid elements in the thymus and peripheral, secondary lymphoid organs of higher vertebrates. Their origin and functional significance are a matter of controversy. In the present investigation we analyze, for the first time, the nature of presumptive interdigitating cells of the thymus of an ectothermic vertebrate, the turtle Mauremys caspica. This model is specially useful because of the seasonal variations that affect the reptilian lymphoid organs. Immature pro-interdigitating cells and phagocytosing mature interdigitating cells are described with special emphasis on their ultrastructural characteristics and possible relationships with monocytes and macrophages.     

Electron-microscopic study of the development of the periarteriolar zone in splenic white pulp of rats

Summary To obtain more information concerning the origin of interdigitating cells, the postnatal development and morphology of the periarteriolar lymphatic sheath in splenic white pulp of rats was investigated by light- and electron-microscopy. Special attention was paid to the ontogeny of interdigitating cells. The spleens of the animals were studied in the age range from 1 h to 28 days after birth.The splenic white pulp of neonatal rats consists only of a few reticuloblasts, which are concentrically arranged around central arterioles. After 21 h an increase in promonocytes and monocytes was noted. Between the fifth and seventh postnatal day monocytogenic cells with a light and almost translucent cytoplasm appear, which display long cytoplasmic projections between the adjacent cells. Neighbouring lymphocytes often insert finger-like processes into the invaginated cellular membrane of these transitional forms. This intimate cellular contact is supported by zonulae occludentes. These cells represent transitional forms between monocytes and interdigitating cells.From seven days of age onwards typical interdigitating cells were present as in adult animals. After the differentiation into an inner and outer periarteriolar lymphatic sheath, the T-cell-dependent area of splenic white pulp has attained its adult appearance and further changes are not to be expected.On the basis of these findings, it is highly probable that interdigitating cells develop via transformation of monocytes.This investigation was supported by the Deutsche Forschungsgemeinschaft (^*He 537)     

The effect of the seasonal cycle on the splenic leukocyte functions in the turtle Mauremys caspica

The reptile immune system is strikingly affected by seasonal variations, which induce changes in the structure of the lymphoid organs and in the function of the leukocytes. The aim of this work is to study several functions of splenic leukocytes from the turtle Mauremys caspica along its seasonal cycle. The functions assayed were adherence to substrate, mobility directed to a chemoattractant gradient (chemotaxis), lymphoproliferative response to mitogens, antibody-dependent cellular cytotoxicity, and natural killer-like cell-mediated cytotoxicity. Splenic leukocytes showed a positive response in all the assays, and this response was similar to that of mammals. In regard to the effect of the seasonal cycle, we have observed in winter a low adherence to substratum and high chemotaxis and cytotoxic activity, whereas in spring, only lymphoproliferation induced by mitogens showed high values except with lipopolysaccharide, which did not induce any seasonal variation in proliferation percentages. In summer, a high chemotaxis and cytotoxicity were observed, while in autumn, adherence to substratum was increased, but chemotaxis, cytotoxicity, and proliferation were clearly diminished. Our results demonstrate that splenic leukocyte functions are affected by the seasonal cycle, which induces a different pattern of response depending on the function studied.     

In vitro proliferation of blood,spleen and thymus leukocytes from the turtle Mauremys caspica

Cell-mediated immune responses is commonly evaluated by cell proliferation assays. Mitogens are known to induce a vigorous proliferative response in lymphoid cells from mammals but relatively fewer studies have investigated mitogen-mediated lymphoproliferation in non-mammalian animals. In the present work, we incubated spleen, thymus and blood leukocytes with phytohaemagglutinin (PHA), concanavalin A (Con A), lipopolysaccharide (LPS) and pokeweed mitogen (PWM), by different times of incubation (96 and 120 h) and at different concentrations. Our results show that the optimal mitogen concentrations inducing proliferation on leukocytes from Mauremys caspica were 20 microg/ml PHA, 1 microg/ml Con A, 12.5 microg/ml LPS and 1/150 dilution PWM. The optimal time of incubation was dependent on the type of leukocytes (peripheral blood leukocytes, splenic leukocytes or thymic cells) and the mitogen utilized.     

The immune response of thymic cells from the turtle Mauremys caspica

In the present work several known mammalian leukocyte functions such as directed mobility, proliferative response to mitogens, antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK)-mediated cytotoxicity were studied in female and male Mauremys caspica turtles. Since the reptilian thymus shows seasonal variations in its structure, we have performed all the assays along the seasonal cycle. Our results show that thymic cells from M. caspica are able to migrate through a chemo-attractant gradient, to proliferate in response to the mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A), and to kill tumoral target cells by both ADCC-mediated and NK-mediated cytotoxicity. Those functions were differentially affected by the seasonal cycle; in general, in autumn the functions studied showed the smallest values for both sexes, while in summer the highest values of cytotoxicity and chemotaxis were found in females. The proliferative responses to PHA and Con A were higher for both sexes in spring and for females in winter than in the other seasons. In summary, thymic cells from M. caspica show a wide range of immune functions, and these are modulated heterogeneously by the seasonal cycle in both sexes.     

Ventilatory responses to temperature variation in the fresh water turtle,Mauremys caspica leprosa

A study of lung gas exchange in the fresh water turtle Mauremys caspica leprosa at normal physiological body temperatures (15, 25 and 35 °C) was extended to extreme temperatures (5 and 40 °C) to determine whether the direct relationship between body temperature and ventilatory response found in many lung-breathing ectotherms including other chelonian species was maintained. From 5 to 35 °C the lung ventilation per unit of O₂ uptake and CO₂ removed declined with temperature. Consequently, lung CO₂ partial pressure increased with temperature. Its value was maintained within narrow limits at each thermal constant, suggesting a suitable control throughout the complete ventilatory cycle. At 40 °C the ventilatory response showed the opposite trend. The ratios of ventilation to lung gas exchange increased compared to their values at 35 °C. The impact of this increased breathing-lowering the estimated mean alveolar CO₂ partial pressure-was nevertheless less than expected due to an increase in calculated physiological dead space. This suggests that the relative hyperventilation in response to hyperthermia found in Mauremys caspica leprosa is related to evaporative heat loss.Abbreviations BTPS body temperature, ambient pressure, saturated with water vapour - CTM critical thermal maximum - F_N2 fractional concentration of nitrogen - PA _CO2or PL _CO2 alveolar or lung CO₂ pressure - PA_O2or PL_O2 alveolar or lung O₂ pressure - PI_O2 inspired O₂ pressure - R respiratory exchange ratio - STPD standard temperature, standard pressure, dry - T _a ambient temperature - T _b body temperature - VA alveolar ventilation - VA/VCO₂ relative alveolar ventilation (alveolar ventilation per unit of CO₂ removed) - VO₂ O₂ uptake - VCO₂ CO₂ output - V _D anatomical dead space volume - V _D physiological dead space volume - VE/VO₂ ventilatory equivalent for O₂ - VE pulmonary ventilation or expiratory minute volume - VE/VCO₂ ventilatory equivalent for CO₂ - V _T tidal volume     

Interdigitating cells in the white pulp of the human spleen

Summary Interdigitating cells are demonstrated as a special type of fixed cell in the periarteriolar lymphocytic sheaths of the human spleen. These cells show typical ultrastructural features as well as a characteristic enzyme histochemical pattern that distinguish them from other reticular cells in the splenic white pulp.     

Ultrastructure of the jugular body of Rana pipiens

Summary The jugular bodies in adult Rana pipiens, are surrounded by a capsule of mesothelium and connective tissue, and their parenchyma consists of cell cords arranged in a sinusoidal network. The cell cords are formed by irregular reticular cells, showing numerous filaments and joined together by zonulae adhaerents. The intercellular spaces are filled by reticular fibres and free cells. These latter are small and medium lymphocytes, lymphoblasts, and developing and mature plasma cells. Additionally, free macrophages, neutrophils and acidophils also occur. Sinusoidal blood vessels show thin walls with numerous filaments and pinocytotic vesicles. They exhibit a discontinuous basement membrane, and tight junctions frequently occur between endothelial cells. Occasionally, lymphatic vessels are found and the innervation is principally vasomotor, although nerve endings appear remarkably near reticular cells and lymphocytes. The jugular bodies of adult R. pipiens are plasma cell and antibody-forming organs, whose functional significance is discussed in relation to their ultrastructural organization.     

Seasonal variations in the activity in vitro of peripheral blood granulocytes in the turtle Mauremys caspica

• 1.1. In the present work we have studied different aspects of the phagocytic process in peripheral blood granulocytes from the turtle Mauremys caspica: (a) tissue adherence capacity, (b) spontaneous mobility and chemotaxis, (c) attachment and ingestion of foreign cells (Candida albicans) or inert particles (latex beads), and (d) capacity to digest ingested material measured by nitroblue tetrazolium (NBT) reduction. These studies were carried out in vitro at 37°C in autumn, winter, spring and summer.
• 2.2. The adherence index showed significantly higher values in autumn, and smaller values in winter. The spontaneous mobility was not affected seasonally, but the chemotaxis was significantly increased in winter and decreased in autumn. The numbers of C. albicans attached and ingested as well as latex beads ingested per 100 granulocytes were higher in autumn and summer, and lower in winter and spring. The digestion capacity was greatly decreased in summer.
• 3.3. The comparison between the results obtained here and those found using the same techniques in blood human granulocytes showed that these animals are endowed with an adequate phagocytic response.

     

Ultrastructural changes in the thymus of the turtle Mauremys caspica in relation to the seasonal cycle

Summary Changes in the ultrastructure of the thymus of the turtle Mauremys caspica, with special reference to its non-lymphoid components, were studied in relation to the seasonal cycle. The thymic cortex contains framework-forming epithelial-reticular cells and free macrophages, while the medulla includes, in addition, mature and presumptive pro-interdigitating cells. The ultrastructural features of these cells are generally similar to those described for non-lymphoid components of the mammalian thymus. The turtle thymus undergoes cortical involution in spring, with recovery periods in May–June and during autumn. A moderate involution occurs in winter. At the beginning of spring, cortical (but not medullary) epithelial-reticular cells show degenerative changes, probably related to high levels of circulating testosterone. In spring and autumn, mature interdigitating cells are absent, but macrophages, monocytes, and pro-interdigitating cells are found. During May–June, the cortical epithelial-reticular population recovers and macrophages, monocytes, and interdigitating cells are actively phagocytic. In summer, the epithelial-reticular cells in both cortex and medulla display normal ultrastructural features; mature and immature interdigitating cells are absent and some macrophages are detected occasionally. The results suggest that non-lymphoid components of the reptilian thymus can play a role in governing T-lymphocyte differentiation, and that the thymic cortex and medulla exhibit different cycles of seasonal activity.     

Specific localization of epidermal-type fatty acid binding protein in dendritic cells of splenic white pulp

Dendritic cells in the splenic white pulp of mice were intensely immunoreactive for epidermal-type fatty acid binding protein (E-FABP). This specific immunostaining revealed a clear difference in morphology between the dendritic cells in the periarterial lymphoid sheath (PALS) and follicular dendritic cells in the follicles in terms of cell sizes and process branching. No immunoreactivity was detected in dendritic cells in the marginal zones and the red pulp, although endothelial cells of almost all capillaries in the red pulp were immunoreactive for E-FABP. After peritoneal injection of lipopolysaccharide, the immunoreactive cells in PALS progressively enlarged and became rounded in shape with a peak in size at 24 h postinjection and they eventually resumed the dendritic form at 48 h postinjection. Within each of the enlarged immunoreactive cell perikarya were included small immunonegative apoptotic cells, presumptive lymphocytes. Taken together, E-FABP is useful as a marker for dendritic cells in the splenic white pulp, and may be involved through combination with fatty acids in antigen presentation and retention as well as in cytokine production.     

Immunocytochemical study of the hypothalamic magnocellular neurosecretory nuclei of the snake Natrix maura and the turtle Mauremys caspica

Summary An immunocytochemical study of the magnocellular neurosecretory nuclei was performed in the snake Natrix maura and the turtle Mauremys caspica by use of antisera against: (1) a mixture of both bovine neurophysins, (2) bovine oxytocin-neurophysin, (3) arginine vasotocin, and (4) mesotocin. Arginine vasotocin- and mesotocin-immunoreactivities were localized in individual neurons of the supraoptic and paraventricular nuclei, with a distinct pattern of distribution in both species. The same cells appeared to be stained by the anti-oxytocin-neurophysin and anti-mesotocin sera. The supraoptic nucleus can be subdivided into rostral medial and caudal portions. In N. maura, but not in M. caspica, neurophysin-immunoreactive neurons were found in the retrochiasmatic nucleus. No immunoreactive elements were seen in the suprachiasmatic nucleus of both species after the use of any of the antisera. A dorsolateral aggregation of neurophysin-containing cells, localized over the lateral forebrain bundle, was present in both species. Magnocellular and parvocellular neurophysin-immunoreactive neurons were present in the paraventricular nucleus of both species. In the turtle, the paraventricular neurons were arranged into four distinct layers parallel to the ependyma; these neurons were bipolar with the major axis perpendicular to the ventricle, and many of them projected processes toward the cerebrospinal-fluid compartment. In N. maura a group of large neurons of the paraventricular nucleus was found in a very lateral position. The posterior lobe of the hypophysis and the external zone of the median eminence contained arginine vasotocin- and mesotocin-immunoreactive nerve fibers. The lamina terminalis of both species was supplied with a dense bundle of fibers containing immunoreactive neurophysin. Neurophysin-immunore-active fibers were also present in the septum, some telencephalic regions, including the cortex and the olfactory tubercule, in the paraventricular organ, and the periventricular and periaqueductal gray of the brainstem.This work was partially supported by a Grant S-85-39 from the Direccion de Investigaciones, Universidad Austral de Chile to E.M. Rodriguez     

Apical membrane potentials and intracellular potassium and sodium activities in the gallbladder of the freshwater turtle Mauremys caspica

Open-tip and liquid ion-exchanger microelectrodes were used to measure transapical membrane potential (Va), fractional voltage ratio (fa) and intracellular sodium and potassium activities (aiNa, aiK) in Mauremys caspica gallbladder under open circuit conditions. The average values (+/- SEM) for Va and fa were -32 +/- 3 and 0.20 +/- 0.03 mV respectively. aiNa and aiK were, respectively, 17 +/- 4 and 82 +/- 7 mM. These results suggest that the mechanisms of Na+ and K+ transport in this tissue are essentially similar to those observed in leaky epithelia in general.     

中华鳖精子的超微结构

爬行纲在动物进化中占有重要地位,而且爬行纲动物种类繁多,生态环境和生活习性差异颇为明显。关于爬行动物生殖特性的资料很有限,而且由于动物种类的复杂性,不同动物的研究结果差异较大(Gustavo et al.,2004),缺乏统一的结构模式。精子的形态结构与动物的生殖特性直接相关,在一     

斑马鱼脾显微与超微结构

应用光学显微镜和透射电镜对斑马鱼(Daniorerio)脾的显微与超微结构进行了观察。光镜结果表明,斑马鱼的脾主要由脾髓和网状组织两部分构成,脾髓分布于整个脾,分为白髓和红髓,但界限不明显,呈混合分布。红髓染色相对较浅,占脾实质的大部分区域,主要由密集的红细胞构成,可见脾窦结构,脾索结构不明显。白髓染色较深,主要由密集的淋巴细胞构成,淋巴细胞的胞核染色较深,呈深紫色。经Gordon-Sweet银染显示网状纤维后,可清晰观察到脾小结,从而可清晰区分红髓与白髓,脾小结与哺乳动物的类似,主要是由密集的淋巴细胞构成。酸性磷酸酶染色可见斑马鱼脾内有大量吞噬性细胞存在。电镜结果显示,红髓分布有不同类型的红细胞、血小板和浆细胞,白髓分布有淋巴细胞、巨噬细胞、单核细胞、嗜中性粒细胞、浆细胞和网状细胞,淋巴细胞胞质内含有多泡体,这可能与淋巴细胞发挥免疫功能有着密切联系。斑马鱼脾与大多数硬骨鱼相似,未观察到椭球这一特殊结构。     

Ultrastructure of lymphatic capillaries in the human dental pulp

Summary Occlusal intradentinal cavities, prepared in normal human premolars and third molars to be extracted for orthodontic reasons, were filled for 7 to 11 days with gutta percha. A superficial pulpitis with localized small abscesses developed in the pulp chamber. Under local anesthesia, 0.2 to 0.3 cc of sterile colloidal carbon was injected in the pulp horn and the teeth were extracted 1 to 3 h later. Lymphatic capillaries could thus be identified in the pulpal tissues. They were characterized by a thin endothelium with occasional large intercellular clefts, absence or incompleteness of basement membrane, absence of pericytes, absence of luminal red blood cells, and presence of a filamentous material between the endothelium and the surrounding collagen fibrils. Moreover, some structural variations were observed.     

The significance of the subcompartments of the marginal zone for directing lymphocyte traffic within the splenic pulp of the rat

Summary These studies were designed to gain more detailed information on the sites of lymphocyte migration into the marginal zone, on lymphocyte segregation within this area and on subsequent migration of the cells in individual compartments of the rat spleen. A lymphocyte population enriched in T-lymphocytes was obtained from rat lymph nodes and was labeled with 5-(³H)uridine in vitro. Observations on localization of labeled lymphocytes at short time intervals following their intravenous transfusion into syngeneic recipients, indicate that the sites of emigration from the blood are the internal and external layers of the marginal zone. From here, the cells migrate towards the periarteriolar lymphoid sheath and the red pulp.Fellow of the Alexander-von-Humboldt-Stiftung. On leave from the Department of Histology and Embryology, Institute of Biostructure, Academy of Medicine, ul. Swiecickiego 6, PL-60-781 Pozna, Poland